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谷氨酸棒杆菌L-天冬氨酸α-脱羧酶基因的克隆及重组酶性质研究

, PP. 110-115

Keywords: L-天冬氨酸α-脱羧酶,谷氨酸棒杆菌,表达,纯化,酶学性质

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Abstract:

从谷氨酸棒杆菌(Corynebacteriumglutamicum)中克隆得到L-天冬氨酸α-脱羧酶基因panD。将该基因连接到pET-24a载体上,并在EscherichiacoliBL21(DE3)中实现了成功表达。蛋白电泳检测显示,PanD自裂解后形成的α亚基和β亚基,即PanD重组酶具有自身加工功能。重组酶的比酶活高达2.60U/mg(156mmol/g?h)。酶学性质研究表明,其最适温度为55℃,最适pH为7.0。80℃条件下,PanD的半衰期约为40min。在37℃,pH7.5条件下,Km值为4.26mmol/L,Vmax为35.97nmol/min,kcat为1.02/s,催化效率kcat/Km为0.24L/mmol?s。

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